One super-resolution optical microscopy technique, called super-resolution fluorescence optical microscopy, consists in measuring the position and the physical characteristics of fluorophores incorporated into the observed medium.
The fluorophores are excitable nanoscopic light emitters that, when they are excited by incident light, emit a certain number of photons before self-destructing. The objective of super-resolution optical methods is to capture these photons so as to locate and/or characterize the fluorophores before the latter self-destruct.
For example, the PALM (acronym of Photo-Activation Localization Microscopy) and STORM (acronym of Stochastic Optical Reconstruction Microscopy) methods are known super-localization microscopy techniques.
One common point of these techniques is that a small number of fluorophores are activated in the medium so as to be able to greatly zoom in onto a single fluorophore and to determine the center of the focal spot generated by said fluorophore.
These observation techniques thus allow intracellular structures to be observed with a precision of about ten nanometers.
These techniques however have drawbacks.
Thus the precision of the position obtained by measuring the center of the focal spot is limited and dependent on the square root of the number of photons received from the fluorophore. Moreover, the emission originating from a rotationally immobile dipolar emitter, such as certain fluorophores, is anisotropic and dependent on the polarization of the exciting light. This creates uncertainty in the obtained position of the fluorophore. Moreover, the need to zoom in onto the fluorophore in order to view the focal spot substantially limits the size of the field of observation.